CN101186932A - Method for synchronously producing hypocrellin and shiraia bambusicola polysaccharides - Google Patents
Method for synchronously producing hypocrellin and shiraia bambusicola polysaccharides Download PDFInfo
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Abstract
The invention relates to the field of bio-fermentation, in particular to a method of producing hypocrellin pigments and polysaccharides simultaneously with solution fermentation. In the art of producing hypocrellin pigments and polysaccharides simultaneously with solution fermentation of the invention, bottle-shaking culture is firstly proceeded, convolution culture is the second step, the enlarging culture of the shaking bottle is the third step and fermentation culture is the final step. In the step of bottle-shaking culture, strain of the hypocrellin is utilized as parent strain, bacteria on the inclined surface of the tube are introduced to a shaking bottle inside which liquid medium is arranged, and the charging coefficient of the shaking bottle is 0.1 to 0.25. In the step of convolution culture, the rotation speed of the convolution culture is 150 to 250 turnings every minute and the culture temperature is 22 to 27 DEG C. In the third step of enlarging culture of the shaking bottle, the culture material achieved by the bottle-shaking culture is shifted to the culture medium of the enlarging culture of the shaking bottle to proceed culture, the inoculation amount is 2-10%, the rotation speed of a rotation bed is 150-250 turnings every minute, the culture temperature is 22 to 27 DEG C, and the culture period is 72 to 120 hours. In the final step of fermentation culture, the culture material achieved by the enlarging culture of the shaking bottle is transferred to a fermentation pot inside which fermentation culture medium is installed to be cultured, the inoculation amount is 5-10%, and the culture temperature is 22 to 27 DEG C. The hypocrellin fermentation liquid is achieved after the fermentation is finished, and the hypocrellin pigments and polysaccharides are respectively achieved by separating and purifying the achieved fermentation liquid.
Description
Technical field
The present invention relates to the biological fermentation field, particularly relate to the method that a kind of solution fermentation is produced hypocrellin and tabasheer polysaccharide simultaneously.
Background technology
Bamboo parasitic fungus (Shiraia bambusicola Henn.) is called red dumpling, the red dumpling of bamboo, bamboo cocoon, the red spot bacterium of bamboo, light chrysanthemum, nandina, henon bamboo, bamboo etc., parasitizes the sporophore (also whistle seat) that forms on the specific bamboo class for Hypocreaceae (Hypocreaceae) fungi tabasheer.It is warm in nature lightly seasoned, have cough-relieving dispel pain, stimulate the circulation of the blood and cause the muscles and joints to relax, effect that expelling wind and removing dampness, invigorating the spleen and replenishing QI, blood circulation promoting and enriching, the stasis of blood of loosing are stimulated the menstrual flow, cure mainly apoplexy, infantile convulsion, peratodynia etc.Tabasheer is in cold of insufficiency type and stomachache, rheumatic arthritis, the trachitis of being used for the treatment of among the people, and diseases such as Whooping cough, sciatica, wound, anaemia headache are a kind of important natural resources of Chinese medicinal materials of China.
Studies show that the main active ingredient in the bamboo parasitic fungus is hypocrellin, tabasheer polysaccharide, N.F,USP MANNITOL, ergosterol etc.Hypocrellin is present known photosensitizers good in visible region, has significant Phototherapy, has been used for treating tetter clinically, as leukoplakia vulvae and softening keloid.What have more application prospect is the effect that hypocrellin has good photosensitive killing tumor cell and inhibition hiv virus HIV-1, and can be used as novel photo-activation pesticide and potential photoelectric conversion material,
The tabasheer polysaccharide mainly is made up of monose such as L-arabinose, D-semi-lactosi, D-glucose, D-seminoses, can improve immunity, and hepatitis is had certain curative effect.
In recent years, be the bacterium that sets out with bamboo parasitic fungus, the existing research report of fermentative Production hypocrellin, as:
Document: Cai Yujie, Ding Yanrui magnifies soldier etc. bamboo parasitic fungus solid state fermentation hypocrellin Study on Conditions. and biotechnology, 2004,14 (4): 46-47 has reported with solid state fermentation and has utilized bamboo parasitic fungus to produce the condition of hypocrellin;
Document: stone Kweiyang, magnify the soldier, Lou Zhihua etc.Generate the research of hypocrellin under the bamboo parasitic fungus liquid culture condition. the medicine biotechnology, reported the condition of with solution fermentation utilizing bamboo parasitic fungus production hypocrellin at 2004,11 (5): 299～301;
Document: Chen Jiajia, Li Zhaolan, Jiao Qingcai.The liquid-state fermentation technology of the asexual strain of tabasheer. Chinese medicinal materials, 2005,28 (12): 1049-1051 has reported substratum composition and the fermentation condition that utilizes the bamboo parasitic fungus liquid fermenting to produce hypocrellin.
Existing research all only relates to the fermentative Production hypocrellin, because hypocrellin and tabasheer polysaccharide are respectively secondary metabolite and primary metabolite, both fermentative production conditions differ, and need in addition integrated survey to optimize the production of hypocrellin and tabasheer polysaccharide simultaneously.Hypocrellin and tabasheer polysaccharide all are the biologically active substances with applications well prospect, produce hypocrellin simultaneously and the tabasheer polysaccharide can obtain this two kinds of materials simultaneously with solution fermentation, but still do not have at present about produce hypocrellin and tabasheer STUDY ON POLYSACHAROSE report simultaneously with solution fermentation.
Summary of the invention
The purpose of this invention is to provide a kind of technology of producing hypocrellin and tabasheer polysaccharide simultaneously, to obtain hypocrellin and these two kinds of active substances of tabasheer polysaccharide with applications well prospect with solution fermentation.
Liquid fermentation process of the present invention comprises with the bamboo parasitic fungus being starting strain, activate through slant strains, carry out liquid shaking bottle cultivation, liquid shaking bottle enlarged culturing base and fermentation culture and obtain using bamboo parasitic fungus fermentation liquid, from the gained fermented liquid, after separation and purification, obtain hypocrellin and tabasheer polysaccharide respectively.
Particularly, the present invention produces in the technology of hypocrellin and tabasheer polysaccharide simultaneously with solution fermentation, the bacterial classification bamboo parasitic fungus of being adopted (Shiraia bambusicola Henn.) is a kind of known bacterial classification, pick up from the Yongjia County, Zhejiang Province, its isolation identification document such as Wei Jingchao, " fungi identification handbook ", Shanghai science tech publishing house, published p238 in 1979; Liu Tianhui, " edible mushrooms outline ", China prospect press published p26 in 1987; Zhang Hao etc., " hypocrellin produces screening and the evaluation of bacterium ", biotechnology, 2002,12 (4), p19-20, the deposit number that China Committee for Culture Collection of Microorganisms provides at the common micro-organisms center is: CGMCCNo.2201.
The present invention produces in the technology of hypocrellin and tabasheer polysaccharide simultaneously with solution fermentation, described slant strains activation medium is the potato substratum, this substratum all has statement in multiple textbook and paper, for example referring to Zhu Gejian, Wang Zhengxiang, " industrial microorganism experimental technique handbook, Beijing, China Light Industry Press, p367, the application quote the document as a reference.22～27 ℃ of slant strains culture temperature, incubation time 96～240 hours.
The present invention produces in the technology of hypocrellin and tabasheer polysaccharide simultaneously with solution fermentation, carry out shake-flask culture earlier: bamboo parasitic fungus (the Shiraia bambusicola Henn.) bacterial strain that with the deposit number is CGMCC No.2201 is a starting strain, shaking in the bottle of liquid nutrient medium is equipped with in the test tube slant bacterial classification access of 10～100mg dry weight/L, shaking bottled liquid coefficient is 0.1～0.25, the cultivation of circling round, the rotating speed of cultivating that circles round is 150～250 rev/mins, 22～27 ℃ of culture temperature, incubation time 72～120 hours; Shake a bottle enlarged culturing then, shake-flask culture gained culture moved in the substratum shake bottle enlarged culturing cultivate, inoculum size 2～10% (percent by volume), shaking speed are 150～250 rev/mins, 22～27 ℃ of culture temperature, incubation time 72～120 hours; Carry out fermentation culture at last, change in the fermentor tank that fermention medium is housed and cultivate shaking a bottle enlarged culturing gained culture, inoculum size 5～10% (percent by volume), 22～27 ℃ of culture temperature, fermentor tank pressure 0.05MPa, 100～180 rev/mins of mixing speed, ventilation 1:0.3～1v/v/m, incubation time 96～150 hours.Obtain the tabasheer fermented liquid after the fermentation ends, from the gained fermented liquid, after separation and purification, obtain hypocrellin and tabasheer polysaccharide respectively.
The present invention produces in the technology of hypocrellin and tabasheer polysaccharide simultaneously with solution fermentation, shake-flask culture base, the substratum that shakes a bottle enlarged culturing base, seeding tank seed culture and fermentation culture consist of, contain following ingredients (unit: gram): carbon source 20～40, nitrogenous source 2～4, K in each premium on currency
2HPO
40.75～2, KCl 0.5～1.0, MgSO
40.5～1.0, FeSO
40.1～0.5, pH 5.5～6.5.Wherein carbon source comprises glucose, maltose, sucrose or murphy juice etc.; Nitrogenous source comprises yeast extract paste, peptone, urea or (NH
4)
2SO
4Deng.
More specifically, preparation technology of the present invention realizes by following steps:
(1) the bamboo parasitic fungus bacterial classification that takes out above-mentioned preservation inserts in the slant medium of new preparation 22～27 ℃ of culture temperature, incubation time 96～240 hours.The moiety of wherein said slant medium is to contain following ingredients (unit: gram): potato 200 (liquor), glucose 20, agar 20, pH5.5～6.5 in each premium on currency.
(2) shaking in the bottle of shake-flask culture base is equipped with in the access of the slant strains in the step 1, consisting of of shake-flask culture base contains following ingredients (unit: gram): carbon source 20～40, nitrogenous source 2～4, K in each premium on currency
2HPO
40.75～2, KCl 0.5～1.0, MgSO
40.5～1.0, FeSO
40.1～0.5, pH 5.5～6.5; The shake-flask culture condition is: 22～27 ℃ of culture temperature, 150～250 rev/mins of shaking speed, incubation time 72～120 hours.
(3) circle round and cultivate the gained fermenting culture and change over to be equipped with and shake the shaking in the bottle of bottle enlarged culturing base shaking bottle in the step 2, the substratum that shakes bottle enlarged culturing consists of, and contains following ingredients (unit: gram): carbon source 20～40, nitrogenous source 2～4, K in each premium on currency
2HPO
40.75～2, KCl 0.5～1.0, MgSO
40.5～1.0, FeSO
40.1～0.5, pH5.5～6.5; The culture condition that shakes bottle enlarged culturing is: inoculum size 2～10% (percent by volume), 22～27 ℃ of culture temperature, 150～250 rev/mins of shaking speed, incubation time 72～120 hours.
(4) change in the fermentor tank that fermention medium is housed and cultivate shaking a bottle enlarged culturing gained culture in the step 3, the substratum of fermentation culture consists of, and contains following ingredients (unit: gram): carbon source 20～40, nitrogenous source 2～4, K in each premium on currency
2HPO
40.75～2, KCl 0.5～1.0, MgSO
40.5～1.0, FeSO
40.1～0.5, pH5.5～6.5; The culture condition of fermentation culture is: inoculum size 5～10% (percent by volume), fermentation tank pressure 0.05MPa, 22～27 ℃ of culture temperature, 100～180 rev/mins of mixing speed, ventilation 1:0.3～1v/v/m, incubation time 96～150 hours.
(5) with centrifugal 30 minutes of 10000 rev/mins of step 4 gained fermenting cultures, separate bamboo parasitic fungus mycelium and fermentation clear liquid.
(6) gained fermentation clear liquid in the step 5 is evaporated to 1/5th to 1/10th of original volume, adds four times of amount ethanol, 4 ℃ left standstill 24 hours, 5000 rev/mins of centrifugal 20 minutes collecting precipitations, with the throw out water dissolution,, promptly get the tabasheer polysaccharide again through ethanol sedimentation, lyophilize.With step 5 gained bamboo parasitic fungus mycelium through cytoclasis, organic solvent (acetone, chloroform or ethyl acetate etc.) extraction, vacuum concentration, again the concentrated solution lyophilize is obtained hypocrellin.
Can in bamboo parasitic fungus liquid fermenting process, obtain hypocrellin and tabasheer polysaccharide simultaneously by present method with higher productive rate.For present employing liquid fermenting was only produced hypocrellin, present method did not increase extra equipment, produced when realizing hypocrellin and these two kinds of activeconstituentss of tabasheer polysaccharide, can increase substantially the economic benefit of production process.Hypocrellin and tabasheer polysaccharide all have higher pharmaceutical use, can be widely used in industries such as medicine and health care, and therefore, the present invention is a kind of hypocrellin and tabasheer polysaccharide production method with applications well prospect.
Embodiment
In order further to set forth material and implementing process involved in the present invention, provided following embodiment, still, the scope that these embodiment do not limit the present invention in any way.
Embodiment 1
Bacterial classification adopts bamboo parasitic fungus (Shiraia bambusicola Henn.), and deposit number is CGMCCNo.2201, picks up from the Yongjia County, Zhejiang Province, its isolation identification document such as Wei Jingchao, " fungi identification handbook ", Shanghai science tech publishing house published p238 in 1979; Liu Tianhui, " edible mushrooms outline ", China prospect press published p26 in 1987; Zhang Hao etc., " hypocrellin produces screening and the evaluation of bacterium ", biotechnology, 2002,12 (4), p19～20.
1, shake-flask culture inserts the bamboo parasitic fungus bacterial classification of preservation in the slant medium of new preparation, 25 ℃ of culture temperature, and the slant culture based formulas is (unit is a grams per liter): glucose 25, potato 200 and agar 20.After treating that mycelium covers with the inclined-plane, 5 milligrams of bamboo parasitic fungus mycelium are inserted in the 250mL triangular flask that 50mL shake-flask culture base is housed (5 bottles) totally, place 25 ℃, 180 rev/mins constant temperature shaking tables to cultivate 96 hours.Wherein said shake-flask culture based formulas is to contain following ingredients (unit: gram): glucose 30, (NH in each premium on currency
4)
2SO
43, K
2HPO
40.15 KCl 0.5, MgSO
40.5, FeSO
40.1, pH6.0.
2, shaking bottle enlarged culturing inserts the bacterial classification of above-mentioned shake-flask culture in the 500mL triangular flask that 100mL enlarged culturing base is housed (totally 20 bottles) and cultivates, inoculum size is to insert the 10mL shake-flask seed in every 500mL triangular flask, places 25 ℃, 180 rev/mins constant temperature shaking tables to cultivate 96 hours.The wherein said culture medium prescription that shakes bottle enlarged culturing is to contain following ingredients (unit: gram): glucose 30, (NH in each premium on currency
4)
2SO
43, K
2HPO
40.15 KCl 0.5, MgSO
40.5, FeSO
40.1, pH6.0.
3, fermentation culture inserts the above-mentioned bacterial classification 2000mL that shakes bottle enlarged culturing in the 30L fermentor tank that the 21L fermention medium is housed and cultivates.Keep 25 ℃ of fermentor cultivation temperature, tank pressure 0.05MPa, 150 rev/mins of mixing speed, ventilation 1:0.5v/v/m, incubation time 120 hours.The substratum of wherein said fermentation culture consists of, and contains following ingredients (unit: gram): glucose 30, yeast extract paste 3, K in each premium on currency
2HPO
40.15 KCl 0.5, MgSO
40.5, FeSO
40.1, pH6.0.
After the fermentation ends, gather in the crops mycelium and fermented liquid, carry out the content analysis of hypocrellin and tabasheer polysaccharide by the following method:
Carry out centrifugation with 10000 rev/mins and obtain mycelium (dry weight) 8.81 grams per liters and fermented liquid;
Fermented liquid is evaporated to 1/4th of original volume, adds 95% long-pending ethanol of tetraploid, and 4 ℃ left standstill 24 hours, and 5000 rev/mins of centrifugal 20 minutes collecting precipitations promptly get the tabasheer polysaccharide through 75% washing with alcohol secondary again.With dissolved in distilled water, adopting sulfuric acid-phynol method to measure polysaccharide content is 354.52 mg/litre;
45 ℃ of dryings of mycelium, get 100 milligrams, add 6 milliliter of 95% extraction using alcohol twice, each extraction time 12 hours, merge vat liquor, (reference: " tabasheer content of hypocrellin A measuring method " [J] Zhejiang Forestry Institute journals such as vast stretch of wooded country duckweed, 2002.19 (2): 157～160), calculating content is 166.8 mg/litre in 465nm colorimetric estimation hypocrellin content.
Embodiment 2
The bacterial classification of present embodiment and shake-flask culture thereof, the culture condition that shakes bottle enlarged culturing are formed identical with embodiment 1 with substratum.
The bacterial classification 2000mL that fermentation culture will be shaken bottle enlarged culturing inserts in the 30L fermentor tank that the 21L fermention medium is housed and cultivates.Keep 25 ℃ of fermentor cultivation temperature, tank pressure 0.05MPa, 150 rev/mins of mixing speed, ventilation 1:0.5v/v/m, incubation time 120 hours.The substratum of wherein said fermentation culture consists of, and contains following ingredients (unit: gram): glucose 30, peptone 3, K in each premium on currency
2HPO
40.15 KCl 0.5, MgSO
40.5, FeSO
40.1, pH6.0.
After the fermentation ends, results mycelium and fermented liquid carry out hypocrellin and tabasheer content Determination of Polysaccharide, and measuring method is with embodiment 1.As a result, mycelium dry weight is 9.26 grams per liters, and the tabasheer polysaccharide is 283.17 mg/litre, and hypocrellin is 192.5 mg/litre.
Embodiment 3
The bacterial classification of present embodiment and shake-flask culture thereof, the culture condition that shakes bottle enlarged culturing are formed identical with embodiment 1 with substratum.
The bacterial classification 2000mL that fermentation culture will be shaken bottle enlarged culturing inserts in the 30L fermentor tank that the 21L fermention medium is housed and cultivates.Keep 25 ℃ of fermentor cultivation temperature, tank pressure 0.05MPa, 150 rev/mins of mixing speed, ventilation 1:0.5v/v/m, incubation time 120 hours.The substratum of wherein said fermentation culture consists of, and contains following ingredients (unit: gram): glucose 30, (NH in each premium on currency
4)
2SO
43, K
2HPO
40.15 KCl 0.5, MgSO
40.5, FeSO
40.1, pH6.0.
After the fermentation ends, results mycelium and fermented liquid carry out hypocrellin and tabasheer content Determination of Polysaccharide, and measuring method is with embodiment 1.As a result, mycelium dry weight is 7.94 grams per liters, and the tabasheer polysaccharide is 207.62 mg/litre, and hypocrellin is 158.4 mg/litre.
Embodiment 4
The bacterial classification of present embodiment and shake-flask culture thereof, the culture condition that shakes bottle enlarged culturing are formed identical with embodiment 1 with substratum.
The bacterial classification 2000mL that fermentation culture is shaken bottle enlarged culturing inserts in the 30L fermentor tank that the 21L fermention medium is housed and cultivates.Keep 25 ℃ of fermentor cultivation temperature, tank pressure 0.05MPa, 180 rev/mins of mixing speed, ventilation 1:0.6v/v/m, incubation time 120 hours.The substratum of wherein said fermentation culture consists of, and contains following ingredients (unit: gram): sucrose 30, (NH in each premium on currency
4)
2SO
43, K
2HPO
40.15 KCl 0.5, MgSO
40.5, FeSO
40.1, pH6.0.
After the fermentation ends, results mycelium and fermented liquid carry out hypocrellin and tabasheer content Determination of Polysaccharide, and measuring method is with embodiment 1.As a result, mycelium dry weight is 8.22 grams per liters, and the tabasheer polysaccharide is 217.35 mg/litre, and hypocrellin is 178.9 mg/litre.
Embodiment 5
The bacterial classification of present embodiment and shake-flask culture thereof, the culture condition that shakes bottle enlarged culturing are formed identical with embodiment 1 with substratum.
The bacterial classification 2000mL that fermentation culture will be shaken bottle enlarged culturing inserts in the 30L fermentor tank that the 21L fermention medium is housed and cultivates.Keep 25 ℃ of fermentor cultivation temperature, tank pressure 0.05MPa, 180 rev/mins of mixing speed, ventilation 1:0.6v/v/m, incubation time 120 hours.The substratum of wherein said fermentation culture consists of, and contains following ingredients (unit: gram): glucose 30, urea 3, K in each premium on currency
2HPO
40.15 KCl 0.5, MgSO
40.5, FeSO
40.1, pH6.0.
After the fermentation ends, results mycelium and fermented liquid carry out hypocrellin and tabasheer content Determination of Polysaccharide, and measuring method is with embodiment 1.As a result, mycelium dry weight is 8.25 grams per liters, and the tabasheer polysaccharide is 207.94 mg/litre, and hypocrellin is 161.6 mg/litre.
Claims (4)
1. the method that solution fermentation is produced hypocrellin and tabasheer polysaccharide simultaneously is characterized in that comprising the steps:
(1) shake-flask culture: bamboo parasitic fungus (Shiraiabambusicola Henn.) bacterial strain that with the deposit number is CGMCC No.2201 is a starting strain, shaking in the bottle of liquid nutrient medium is equipped with in bacterial classification access in test tube slant, dress liquid coefficient is 0.1～0.25, the cultivation of circling round, the rotating speed of cultivating that circles round is 150～250 rev/mins, 22～27 ℃ of culture temperature, incubation time 72～120 hours;
(2) shake a bottle enlarged culturing: shake-flask culture gained culture is moved in the substratum that shakes bottle enlarged culturing cultivate, inoculum size 2～10% (percent by volume), shaking speed are 150～250 rev/mins, 22～27 ℃ of culture temperature, incubation time 72～120 hours;
(3) fermentation culture: will shake bottle enlarged culturing gained culture and change cultivation in the fermentor tank that fermention medium is housed over to, inoculum size 5～10% (percent by volume), 22～27 ℃ of culture temperature, fermentor tank pressure 0.05MPa, 100～180 rev/mins of mixing speed, ventilation 1:0.3～1v/v/m, incubation time 96～150 hours; Obtain the tabasheer fermented liquid after the fermentation ends, from the gained fermented liquid, after separation and purification, obtain hypocrellin and tabasheer polysaccharide respectively.
2. the method for claim 1 is characterized in that, the shake-flask culture base, shakes consisting of of a bottle enlarged culturing base, fermention medium, contains following ingredients (unit: gram): carbon source 20～40, nitrogenous source 2～4, K in each premium on currency
2HPO
40.75～2, KCl 0.5～1.0, MgSO
40.5～1.0, FeSO
40.1～0.5, pH5.5～6.5.
3. as claim 1 and the described method of claim 2, it is characterized in that shake-flask culture base, the carbon source of shaking a bottle enlarged culturing base, fermention medium comprise glucose, maltose, sucrose or murphy juice etc.; Nitrogenous source comprises yeast extract paste, peptone, urea or (NH
4)
2SO
4Deng.
4. the method for claim 1 is characterized in that, the purification procedures of described fermented liquid comprises the centrifugation of fermented liquid, obtains supernatant liquor and thalline respectively; Gained supernatant concentration to 1/6～1/12 volume with ethanol sedimentation, with the throw out water dissolution, again through ethanol sedimentation, lyophilize, gets the tabasheer polysaccharide; The gained thalline is through cytoclasis, organic solvent (acetone, chloroform or ethyl acetate etc.) extraction, vacuum concentration, obtain hypocrellin with the concentrated solution lyophilize again.
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| CN102465154A (en) * | 2010-11-04 | 2012-05-23 | 苏州大学 | Method for improving hypocrellin yield in shiraia bambusicola |
| CN101875905B (en) * | 2009-11-06 | 2012-07-25 | 首都师范大学 | Mao bamboo seed endophytic fungi strain for efficiently producing hypocrellin and application thereof |
| CN102690146A (en) * | 2012-05-29 | 2012-09-26 | 浙江农林大学 | Formula and preparation process of medium for culturing medical shiraia bambusicola henn |
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| CN113149819A (en) * | 2021-05-18 | 2021-07-23 | 山东国力生物科技有限公司 | Method for extracting hypocrellin from solid-state fermentation material of tabasheer fungus |
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| CN1421521A (en) * | 2001-11-28 | 2003-06-04 | 刘为忠 | Liquid fermentation process of fungus producing perylene quinone compound |
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| CN102465154A (en) * | 2010-11-04 | 2012-05-23 | 苏州大学 | Method for improving hypocrellin yield in shiraia bambusicola |
| CN102465154B (en) * | 2010-11-04 | 2013-12-04 | 苏州大学 | Method for improving hypocrellin yield in shiraia bambusicola |
| CN102690146A (en) * | 2012-05-29 | 2012-09-26 | 浙江农林大学 | Formula and preparation process of medium for culturing medical shiraia bambusicola henn |
| CN102690146B (en) * | 2012-05-29 | 2016-05-25 | 浙江农林大学 | A kind of culture medium prescription and preparation technology who cultivates medicinal fungi tabasheer |
| CN103169735A (en) * | 2013-03-15 | 2013-06-26 | 江西众心堂制药有限公司 | Traditional Chinese medicine for treating rheumatic arthritis and rheumatoid arthritis |
| CN106916861A (en) * | 2016-11-25 | 2017-07-04 | 温州大学 | It is a kind of while the method for producing Auricularia polysaccharide and melanin |
| CN107099489B (en) * | 2017-06-30 | 2020-10-09 | 苏州大学 | Associated bacterial strain for improving hypocrellin fermentation yield and application thereof |
| CN107099489A (en) * | 2017-06-30 | 2017-08-29 | 苏州大学 | One plant raising hypocrellin fermentation production rate associated bacteria bacterial strain and its application |
| CN110172409A (en) * | 2019-04-26 | 2019-08-27 | 华南理工大学 | One plant height produces tabasheer bacterial strain and its application of hypocrellin A |
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| CN111500657A (en) * | 2020-05-07 | 2020-08-07 | 杭州巴洛特生物科技有限公司 | Method for co-producing hypocrellin A and bamboo yellow exopolysaccharide |
| CN111500657B (en) * | 2020-05-07 | 2023-10-13 | 杭州巴洛特生物科技有限公司 | Method for co-producing hypocrellin A and tabasheer exopolysaccharide |
| CN113149819A (en) * | 2021-05-18 | 2021-07-23 | 山东国力生物科技有限公司 | Method for extracting hypocrellin from solid-state fermentation material of tabasheer fungus |
| CN113233970A (en) * | 2021-05-18 | 2021-08-10 | 山东国力生物技术研究院 | Preparation method of high-purity hypocrellin |
| CN113233970B (en) * | 2021-05-18 | 2022-09-02 | 山东元核生物工程有限公司 | Preparation method of high-purity hypocrellin |
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